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. 2023 Apr 3;75(3):570–584. doi: 10.1007/s43440-023-00480-6

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© The Author(s) 2023, corrected publication 2023

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — Macrophage-derived argininosuccinate bypasses ADI-PEG20 cytotoxicity in MPM cells. A Mean [arginine] and [argininosuccinate] in the supernatant from macrophage and MSTO tumor cells alone and co-cultured (with and without direct cell contact) by 48 h following ADI-PEG20 treatment using LC/MS (n = 3). Bars show mean and standard deviation: *p < 0.05, **p < 0.01, ****p < 0.0001; two-way ANOVA with post hoc Tukey’s test ([arginine], F_7,14 = 92.90, df = 7; p < 0.0001), [argininosuccinate], (F_7,14 = 213.3, df = 7; p < 0.0001); (B Argininosuccinate rescue (using 0.5 µg/ml, i.e., peak concentration measured above) with MSTO tumor cell viability assessed at 2 days in the presence of ADI-PEG20 (n = 6). Bars represent mean values and standard deviation: **p < 0.01; one-way ANOVA with uncorrected Fisher’s LSD post hoc test (F_2,15 = 6.670, df = 2; p < 0.0085); and C the effect of ASL mRNA knockdown in the MSTO MPM cell line on the metabolic resistance conferred by macrophage-derived argininosuccinate. Cell viability was assessed at 4 days by flow cytometry. Bar values representing the mean and standard deviation (n = 3): *p < 0.05, **p < 0.01, ****p < 0.0001; two-way ANOVA with post hoc Tukey’s test (F_9,18 = 17.30, df = 9; p < 0.0001)